To observe and measure ADA turnover kinetics in the absence of an inhibitor. This work will be the basis of our comparison to ADA-AuNP turnover studies.
- Make a 40μM solution of adenosine in buffer (50mM phosphate buffer, pH 7.4)
- Go to Dr. Hartings lab for enzyme kinetics measurements.
- Add 3mL of adenosine solution to the cuvette
- Start your kinetics measurement
- 1ms integration (on front panel)
- 10 scan average (on front panel)
- Set "Save the first available scan every" to 15 seconds (after clicking File>Save)
- Set "Stop after this amount of time" to 10 minutes (after clicking File>Save)
- Set "File Type" to Tab Delimited
- Give the files a directory and a name
- Click accept
- Just before 1 minute add 30ul of 0.01u/mL ADA
- 0.7318 g of NaH2PO4·H2O and 5.2520g Na2HPO4·7H2O in 500mL water --> 50mM Phosphate buffer pH 74
- Adenosine deaminase (ADA) stock
- 1.1mg (24.0units in 1 mg) in 25mL of buffer --> 1.1 units/mL ADA
- ADA for experiments
- 100uL of stock ADA and 900uL buffer --> 0.11 units/mL ADA
The following graph shows the changes of concentrations of Adenosine and Inosine as a function of time.
This area is for any observations or conclusions that you would like to note.
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